The aim of this study was to compare mineral, amino acid composition, physical properties and use in gluten-free bread of seeds of holy basil (Ocimum tenuiflorum) with those of chia seeds (Salvia hispanica). Holy basil and chia seeds have variable chemical compositions. Holy basil is a valuable source of calcium, manganese, and iron andcontains significantly more methionine sulfone and tryptophan. However, higher levels of asparagine, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, leucine, phenylalanine, and lysine were found in chia seeds compared to holy basil seeds. With regard to energy consumption during grinding, holy basil appears to be more cost-efficient. The addition of both whole and ground O. tenuiflorum and S. hispanica had a favorable effect on bread crumb texture. Holy basil and chia swell intensely in water and can be used as hydrocolloid replacements in gluten-free bread.